Method for treating viral infection using topical administration

ABSTRACT

Methods of treating systemic viral infections are disclosed comprising the parenteral administration of pharmacologically effective amounts of ozonides of terpenes in pharmaceutically acceptable carriers. Methods for treating viral lesions are also disclosed. In particular, a method for treating retroviral infections is disclosed. More particularly, a method for treating HIV infections is disclosed. In addition, methods for treating infections of non-retroviral viruses are disclosed. Further, methods for treating T-cell deficiencies are also disclosed. Moreover, a method of producing blood for medical products which is free of viral activity is disclosed.

CROSS REFERENCE TO RELATED APPLICATION

This application is a division of application Ser. No. 363,628, filedJune 8, 1989, now U.S. Pat. No. 4,983,637, which is acontinuation-in-part of U.S. application Ser. No. 211,378, filed June24, 1988 and now abandoned.

This invention relates to methods of medical treatment. Moreparticularly, it relates to the use of ozonides of terpene hydrocarbonsin the treatment of viral infections and certain immune disorders.

BACKGROUND OF THE INVENTION

Methods of medical treatment employing ozonides of oil-soluble compoundsare known in the art, being disclosed, for example, in U.S. Pat. No.925,590 to Neel, U.S. Pat. No. 2,083,572 to McKee, and U.S. Pat. No.4,451,480 to De Villez.

The prior art does not disclose the use of ozonized compounds as anantiviral or immunotherapeutic agent. However, particular types ofozonide structures have been disclosed to have certain pharmacologicalactivity. In U.S. Pat. No. 925,590, Neel reports the use of ozonides ofterpenes and other ozonides for inhalation therapy, because it wasbelieved to have a therapeutic effect for consumption and asthma.Although the Neel patent application was filed in 1902, there haveapparently been no supporting data reported in the intervening yearsthat corroborate the utility theorized by Neel.

Knox, U.S. Pat. No. 1,210,949 discloses use of ozonized castor oil as alaxative. Ozonation of the oil was believed to reduce its toxicity andcreate a germicidal effect.

Johnson, U.S. Pat. No. 2,356,062 discloses the use of ozonides ofglycerine trioleates for external application, because it was believedthat those particular triglycerides had a germicidal, fungicidal anddeodorizing effect.

De Villez, U.S. Pat. Nos. 4,451,480 and 4,591,602, discloses use ofozonides of certain fatty acids, including olive oil, sesame oil, jojobaoil, castor oil and peanut oil, for external use as antimicrobialagents, particularly in the treatment of acne. It is believed that atleast some of these compounds cause unacceptable skin irritation.

So far as can be determined, none of the medical uses of ozonidesdescribed in the prior art have ever been commercialized. Presumably,this lack of commercialization is due to unacceptable side-effects,toxicity, difficulties in storage, or minimal effectiveness. Many ofthese various compositions decompose on standing. Also, to the extentthat the mechanism of action of these compositions can be attributed totheir oxygen content, most of the ozonides known in the prior art havebeen suboptimal because these compounds typically release no more thanabout 18% of their weight as oxygen.

Methods of medical treatment employing antiviral compounds are known inthe art. Most of the research in this area has focused on nucleosideanalogues. Dideoxynucleosides are antiviral nucleoside analogues whichare useful in treating retroviral infections where viral replicationrequires the transcription of viral RNA into DNA by viral reversetranscriptase. Other nucleoside analogues include deoxynucleosides andnucleoside analogues, such as acyclovir and gancyclovir, which have onlya fragment of ribose or other pentose connected to the base molecule.Nucleoside analogues have been shown to be only minimally effective inthe treatment of viral infections that are not caused by retroviruses.

Antiviral agents other than nucleoside analogues are also known. Forexample, amantadine is an antiviral agent that prevents binding ofcertain viruses with their receptor on the cell surface. However,amantadine is ineffective against many known viruses.

Acquired immunodeficiency syndrome (AIDS) is a fatal condition caused bythe human immunodeficiency virus (HIV), a retrovirus. Since AIDS wasidentified as a medical condition in 1981, over 100,000 cases have beenreported worldwide, with over half of these cases in the United States.It is believed that over 2,000,000 people worldwide are carriers of theHIV virus, with infections continuing to spread. Researchers now believethat most of these carriers will one day develop symptoms of AIDS. Noeffective cure is available for AIDS, although dideoxynucleosides andtheir analogues have been shown to prolong life and to reduce theincidence of certain fatal infections associated with AIDS. Among thedideoxynucleoside analogues, AZT has shown the most promise as atreatment for AIDS. However, treatment of AIDS patients with AZT hasproven to be of only poor to moderate effectiveness, and AZT does notcure AIDS. Moreover, in a recent human trial, serious toxicity wasnoted, evidenced by anemia (24%) and granulocytopenia (16%). Clearly,there is a tremendous need for a non-toxic and effective treatment forHIV infection.

It is believed that HIV causes AIDS, in part, by infectinghelper/inducer T4-cells and causing a T4-cell deficiency. Otherconditions may also cause this deficiency, including immunosuppressivetherapy for transplant patients, radio-therapy or chemotherapy in cancerpatients, and congenital immunodeficiencies. Current immune boostingtherapies, such as the use of interleukin-2 or γ-interferon are still inthe experimental stages, and have not yet been proven effective. Noproven effective treatments are currently in use for restoring a normallevel of T-cells. Thus, a need exists for such a treatment.

Transmission of HIV through blood products has been shown to occur. Thediscovery of the HIV antibody test, and its application to bloodproducts prior to release has reduced the incidence of transmissionthrough blood products. However, the HIV antibody test is not 100%effective in detecting the presence of HIV virus particles, in part,because an infected individual may not produce antibodies to HIV for sixmonths or longer after infection. There is, therefore, still a lowincidence of blood products tainted with HIV being released for medicaluse. Moreover, blood products may be tainted with other viruses capableof being transmitted through the blood, such as Hepatitis B. A method oftreating blood products to eliminate viral activity without affectingtheir efficacy in treatments is highly desireable.

Retroviruses other than HIV are known. These include the herpes familyof viruses, HTLV I, and cytomegalovirus (CMV). Infections of theseviruses have been notoriously difficult to treat. No vaccines are knownfor these infections. Although acyclovir has been used in the treatmentof Herpes lesions, toxic side effects are known, and such treatment isnot always effective. Thus, a need exists for non-toxic and effectivetreatments.

Human papilloma viruses are nonretroviral viruses responsible for wartsof the skin or mucous membranes. Common warts are found in as many as25% of some groups, and are most prevalent among children Moreover, theincidence of venereal warts (condylomata acuminata and molluscumcontagiosum) has risen dramatically in the last few years, to the pointthat this condition is one of the most common sexually transmitteddiseases in the United States. Common treatments for warts are oftenpainful and invasive, and involve physical removal of the lesion throughapplication of caustic agents, cryosurgery, electrodessication, surgicalexcision, or ablation with laser. Treatment with nucleoside analogues orinterferon is also sometimes used. However, no treatment of provensafety and efficacy is currently available for warts. Furthermore, atthe present time, no effective methods of prevention are available forwarts other than avoiding contact with infectious lesions. Therefore, aneed exists for a method of treatment and prevention of warts.

Other nonretroviral viruses are responsible for many of the knowninfections in mammals. Vaccines are known for a minority of theseinfections. Measles, rubella, polio, rabies, certain strains ofinfluenza, and mumps are examples of infections caused by viruses forwhich vaccines are known. However, the existence of a vaccine does notlo obviate the need for treatment of individuals already infected. Mostother viruses, including Epstein Barr Virus, and most of theenteroviruses, reoviruses, rhabdoviruses other than rabies, arboviruses,and arenaviruses produce infections for which no vaccines are known.Currently used antiviral treatments for infections of these virusesinclude application of nucleoside analogues or amantadine, and variousinterferon treatments. Unfortunately, use of these treatments is ofminimal or no effectiveness against infections of most of these viruses.The use of currently known antiviral compounds is, at best, moderatelyeffective. Moreover, toxic side-effects are common. Thus a need existsfor a wide-spectrum antiviral agent that is both non-toxic andeffective.

Virtually all humans occasionally suffer from upper respiratoryinfections, such as colds and flu. The symptoms of these infectionsinclude sore throat, runny nose, itchy eyes, and earache. In addition tothese discomforts, the infections are responsible for many days ofabsence from work and contribute to a decrease in worker efficiency.These infections are caused by a wide variety of viruses. Althoughvaccines are known for a minority of flu strains, no effective methodsof prevention are known for most upper respiratory infections, and notruly effective methods of treatment are known for any of theseinfections. A method of treating these symptoms and underlyinginfections would be of tremendous benefit.

Moreover, there are a number of ailments that may or may not be of viralorigin, for which no effective treatments are widely available.Epstein-Barr virus (EBV) is the causative agent in infectiousmononucleosis, and has been implicated in chronic fatigue syndrome. Manyautoimmune disorders, such as systemic lupus erythematosis andrheumatoid arthritis, may be associated with a virus. Whether or notthese diseases are of viral origin, however, they are debilitatingailments for which an effective therapy would be of major importance.

Finally, there are a number of situations, both in research and inmedicine, in which generation of the superoxide radical O₂ ⁻ isadvantageous. Superoxide is commonly generated through the use ofxanthine oxidase acting on xanthine. However, these materials arerelatively expensive and are not particularly suited for many utilities,including in vivo utilities. Thus, a method for generating superoxidethat is safe and inexpensive would be advantageous.

SUMMARY OF THE INVENTION

According to the invention, there are provided novel methods oftreatment and prevention of systemic and local viral infections. Methodsfor treating nonretroviral and retroviral infections in particular arealso provided. More particularly, a method for treating retroviral HIVinfections is provided. Moreover, the invention provides a method oftreating blood products which removes any viral activity present in theblood. Additionally, the invention provides a method of treatingimmunosuppression characterized by T-cell deficiencies.

The invention, in addition, provides pharmaceutical compositions for usein the above novel treatments, containing ozonides of terpenes and apharmaceutically acceptable carrier, and may contain other activeingredients. Preferably, these compositions are in dosage formcomprising a clinically effective amount of the active compound. In onepreferred embodiment of the invention, the pharmaceutical composition iscomprised of an ozonized terpene in a stable injectable composition. Inother preferred embodiments, the pharmaceutical compositions are in theform of nosedrops or nasal sprays, inhalants, throat sprays, eardrops,ophthalmic ointments or drops, vaginal or rectal suppositories, orointments or creams for topical applications.

Moreover, the present invention includes the use of terpene ozonides andother ozonides of unsaturated hydrocarbons to treat autoimmunedisorders, and to produce superoxide radical upon combination with anaqueous system.

DETAILED DESCRIPTION OF THE INVENTION

Terpene hydrocarbons are also known as isoprenoids, because they maygenerally be constructed from isoprene units. Terpene hydrocarbons areusually exact multiples of C₅ H₈. Terpenes are classified according tothe number of isoprene units of which they are composed, as shown inTable 1.

                  TABLE 1                                                         ______________________________________                                               1 hemi-        5 ses-                                                         2 mono-        6 tri-                                                         3 sesqui-      8 tetra-                                                       4 di-          n poly-                                                 ______________________________________                                    

While not limiting the scope of the invention, examples of terpeneswhich may prove especially effective, when used in the method of thepreferred embodiment, include limonene, citronella, alpha-carotene,beta-carotene, Vitamin A, geraniol, linalool, linalyl acetate, andsqualene. Other compounds which are believed to make pharmacologicallyactive terpene ozonides in accordance with the present invention includelimonene, alpha-pinene, loganin, cymene, farnesanes, eudesmanes,acoranes, cedranes, chamigranes, caryophyllanes, illudanes, humulenes,himachalenes, longifolanes, perhydroazulenes, quaianes, quaianolides,and germacranes. Still other compounds which are believed to makepharmacologically active terpene ozonides in accordance with the presentinvention include labdanes, clerodanes, abietic acid, phyllocladene,giberellins, ophiobolin A, retigeranic acid, gasgardic acid, lanosterol,euphol, oleanane, ursane, lupeol, hydroxyhopanone, lupanes, and hopanes.Other particular terpene compounds which are believed to makepharmacologically active terpene ozonides when prepared in accordancewith the present invention include B-selinene, zingibene, camphene,sabinene, ocimene, myrcene, nerol, citral A, citral B, farnesol,bisabolene, phytol, and cecropia hormone. Citral, geraniol, and nerolare particularly preferred terpenes. Ozonides of terpenes have threeoxygen atoms replacing the double bonds at sites of unsaturation,creating a trioxyacyclopentane.

In the preparation of terpene ozonides, the particular desired terpenestarting material is first obtained. A large and representative numberof such terpenes are disclosed in the literature and/or are commerciallyavailable. (Many terpenes are essential oils that have been isolatedfrom various parts of plants or wood by steam distillation orextraction.)

In the ozonide synthesis, ozone is passed through the terpene underconditions that provide for intimate contact between the terpenestarting material and the ozone, such as thin film procedures, sparging,gas entrainment procedures, and the like. On a small scale, for example,the terpene is placed in a vented vessel, and ozone is sparged throughthe material until the reaction is complete. The ozone mayadvantageously be generated with any of the commercially-available ozonegenerators. Such devices include corona discharge tubes through whichoxygen gas may be passed. For example, pure oxygen gas passing throughan ozone generator will typically leave the device as from 2% to 6% O₃(ozone), with the remainder O₂. This ozone mixture may then be spargedthrough the terpene at ambient temperature and pressure until thereaction is complete. Completion may be judged by analyzing the gasexiting the ozonation chamber for ozone. (This may be done by passingthe exit gas through aqueous potassium iodide and determining whetheriodine gas is liberated, or by any other conventional technique.)Alternatively, the reaction may be followed by observing the weight gainof the material undergoing the reaction, by observing changes inphysical characteristics (such as conversion from a liquid form to asoft paste), or by simply calculating the quantity of ozone needed tofully ozonate the material and stopping the reaction when a slightexcess of ozone has passed through the reaction chamber. Because thereaction is exothermic, its progress may also be followed by monitoringthe heat evolved by the reaction medium, and stopping the flow of ozonewhen the mixture ceases to generate heat.

When the termpene is normally a solid, such as β-carotene, it may besolubilized in any suitable saturated nonaqueous solvent system prior toozonation. With all of the terpene ozonides, it is desirable to excludewater lower alcohols, nucleophilic perozides, and proton donors from thereaction mixture and from the final composition in order to preventpremature hydrolysis of the trioxoloane ring.

Other suitable ozonation procedures may be used, such as the proceduresdisclosed in U.S. Pat. Nos. 2,083,572, 3,504,038, and 4,451,480.

In certain preferred embodiments of the present invention, the terpeneozonides are formulated into pharmaceutical preparations. Thesepharmaceutical preparations include one or more terpene ozonides, andmay further include other pharmacautically active ingredients. Inaddition, any of the well-known pharmaceutically-acceptable carriers orexcipients may be combined with termpene ozonides in a well-knownmanner. Suitable diluents include, for example, polyethylene glycol,DMSO, isopropyl myristate, and mineral oil. Conventional coloring,fragrance, and preserving agents may also be provided.

It is believed that the excellent weight to oxygen ratio of some of theterpene ozonides renders them especially effective as antiviral agents.Some of the terpene ozonides are capable of releasing large amounts ofoxygen, up to 30% of the weight of the compound. This is becauseterpenes are highly unsaturated compounds. Ozonation of these compoundsresults in the addition of three oxygen atoms at each site ofunsaturation. In addition, terpene ozonides appear to have significantunexpected pharmacological properties that are different in kind orquality from those of unrelated ozonides disclosed in the prior art.

The toxicity of the terpene ozonides appears to be surprisingly low insystemic use. Our preliminary data

suggest that the LD₅₀ for a representative compound, linalool ozonide,appears to be greater than about 5000 mg/kg in mice. Furthermore, wehave discovered that the irritability of the terpene ozonides issurprisingly low in skin and eye tissues of the rabbit. It is believedthat irritability of the compounds in humans is also surprisingly lowwhen used in accordance with the methods of the preferred embodiments.

These ozonides can be used effectively in the generation of superoxideradical, both in vitro and in vivo. When the ozonide is combined with anaqueous system, gradual decomposition of the ozonide trioxolane ringstructure occurs, with release of superoxide radical. Thus, the presentinvention includes a method for generating superoxide by combining theseozonides with a water-containing system. For example, superoxideproduction is believed to occur when the compounds are administered toan organism, as well as when the compounds are mixed (with or without asurfactant) into a material that contains water. While the inventor doesnot wish to be limited to any particular theory of operation, it isbelieved that at least some of the beneficial and therapeutic propertiesof these ozonides are due to superoxide generation.

We have also discovered that terpene ozonides, injected in suitablepharmacological compositions, are effective for treatment of systemicviral infections. The present invention includes systemic and localizedinjection of terpene ozonides, including intravascular, intramuscular,subcutaneous, intraperitoneal, and other injection techniques. Inaddition, oral administration is also contemplated, preferably in acapsule or other nonaqueous vehicle or system.

In the method of a preferred embodiment, pharmaceutical compositions forsystemic use such as for intravenous, intramuscular, or intraperitonealinjection may contain from about 0.01% to about 99% active ingredient,by weight. More preferred injectable compositions contain from about0.05% to about 45% active ingredient, by weight. Moreover,pharmaceutical compositions for local application in the form ofnosedrops or nasal sprays, inhalants, throat sprays, eardrops,ophthalmic ointments or drops, rectal or vaginal suppositories, orointments or creams for topical applications may contain from about0.01% to 99.9% active ingredient, by weight. More preferred compositionsfor local application contain from about 0.05% to 50% active ingredient,by weight.

Pharmaceutical compositions of preferred embodiments may contain fromabout 0.I% to 99.99% carrier ingredients. The carriers are preferablynon-aqueous, because the presence of water rapidly leads to thedegradation of the pharmacologically active ozonide compounds used inthe preferred embodiment. The carriers employed in pharmaceuticalcompositions for systemic use are, in addition, preferably injectable ororally ingestible. Non-aqueous, injectable carriers for pharmaceuticalcompositions of the preferred embodiment for systemic applicationpreferably include: isopropyl myristate, polyethylene glycol orpolypropylene glycol (in liquid form), and DMSO, more preferablypolyethylene glycol having a molecular weight between about 150 and1500, most preferably about 600. Good results have been realized, forexample, by combining about 4 parts by weight of geraniol ozonide withabout 3 parts by weight polyethylene glycol (m.w. 600). This material isstorage stable, and can be formulated into an injectable material bycombining one part with three parts sterile saline immediately beforeuse. (Although some superoxide production appears to begin immediatelyupon such combination, no significant degradation of the ozonide isbelieved to occur within, say, 5 minutes before the material isinjected.) Another suitable vehicle is epal, comprising roughlyequivalent parts of substantially anhydrous tetradecanol and dodecanol.Non-aqueous carriers suitable for pharmaceutical compositions for localapplication in accordance with the methods of the preferred embodimentinclude: DMSO, hydrogenated vegetable oil, mineral oil, carbomer 934,glycerin, propylene glycol, propyl paraben, polysorbate 60, glycerylstearate, ethanol, and modified food starch.

Therapeutic dosages of the terpene ozonides when used for systemicinjection in accordance with the methods of the preferred embodimentsare preferably in the range of 1 mg to 10 g active ingredient for a 70kg adult one time per day, more preferably in the range of 10 mg to 1 gactive ingredient for a 70 kg adult one time per day, and mostpreferably in the range of 20 mg to 500 mg active ingredient for a 70 kgadult one or two times per day. Therapeutic dosages of the terpeneozonides when used for topical application in the form of creams,ointments, or rectal or vaginal suppositories in accordance with themethods of the preferred embodiments are preferably in the range of 100g to 10 g used one to four times per day for each cm² of affected area,more preferably 1 mg to 1 g used one to four times per day for each cm²of affected area, and most preferably 5 mg to 200 g used one to fourtimes per day for each cm² of affected area. Therapeutic dosages of theterpene ozonides for use in other methods of local application inaccordance with the preferred embodiments, such as nosedrops or nasalsprays, inhalants, throat sprays, eardrops, or ophthalmic ointments ordrops are preferably in the range of 100 g to 1 g per application usedone to four times per day, and more preferably 1 mg to 100 mg perapplication used one to four times per day.

Oral compositions may be given at the same dosage as the injectablecompositions, or may be given at up to twice the injection dosage.

We have discovered that intramuscular injection of a terpene ozonide ina pharmaceutically acceptable carrier, with or without contemporaneousoral administration, is effective in treating the symptoms of AIDS. Apatient receiving this treatment gets fewer of the opportunisticinfections common in AIDS patients. Such a patient also feels lesslethargic and has a generally improved sense of physical well-being.This improvement in symptoms has been shown to be the result of arestoration of normal T4-cell levels after injection with the terpeneozonide. The restoration is believed to be the result of an antiviraleffect of the terpene ozonide.

It is believed that treatment of persons infected with HIV who do notyet express symptoms of AIDS can be effectively treated with systemicinjections of terpene ozonides, for example in the manner of Example 11,in order to prevent the appearance of the symptoms of AIDS.

Furthermore, it is believed that administration of terpene ozonides inaccordance with the present invention will be beneficial in thetreatment of immune disorders other than AIDS. It is believed thatsystemic injection of the terpene ozonides will restore a normal levelof T4-cells in many immunocompromised patients, including patients onimmunosuppressive therapy, chemotherapy, or radio-therapy, and patientswith congenital immunodeficiencies. Lupus and rheumatoid arthritis alsorespond to therapy with the terpene ozonides of the present invention.In a preferred embodiment, restoration of T4-cell levels is accomplishedby systemic injection of terpene ozonides in the manner of Example 11.

It is also believed that treatment of blood products with a terpeneozonide of the present invention prior to its medical use, willeliminate HIV and any other viral activity present in the blood. Fromabout 0.5 to about 10 mg/liter of terpene ozonide can be used intreating blood in, for example, a blood bank.

The terpene ozonides seem to be effective not only against HIVinfection. They also appear to be effective in the treatment of otherretroviral infections, such as Herpes lesions, including chicken pox,EBV infection, or CMV infection. Systemic injection of terpene ozonidesis believed to be effective in treatment of these other retroviralinfections. Additionally, topical application of the terpene ozonides inpharmacologically effective compositions is believed to be effective inthe treatment of lesions of these retroviral infections. Moreover, it isbelieved that the terpene ozonides will be effective against manydisparate viral infections, including viral infections of non-retroviralorigin. In this regard, it is believed that systemic injection of aterpene ozonide in a pharmacologically acceptable carrier or excipientis effective in the treatment of systemic infections caused bynon-retroviral viruses, including Epstein Barr Virus, most of theenteroviruses, reoviruses, rhabdoviruses (including rabies),arboviruses, and arenaviruses. It is also believed that intra-vaginalapplication of a terpene ozonide in a pharmaceutically acceptablecarrier or excipient is effective against condylomata acuminata,molluscum contagiousum, and other viral infections of the vagina. Alsoin this regard, it is believed that topical application of a terpeneozonide in pharmacologically acceptable carrier or excipient isnon-irritating and effective in the treatment of common warts and otherviral lesions of the skin. Further in this regard, it is believed thatapplication of a terpene ozonide in a pharmacologically acceptablecarrier or excipient in the form of nosedrops or nasal sprays,inhalants, throat sprays, eardrops, ophthalmic ointments or drops, iseffective in the treatment of viral infections of the eye, ear, nose,and throat, including upper respiratory infections of viral origin suchas colds and flu. Finally, they appear to be useful in treatment ofrheumatoid arthritis, which may be caused by a viral pathogen, as wellas useful in treatment of other autoimmune disorders.

For example, in treating the common cold, an aerosol mist containing 2ml of the nasal inhalant of Example 10 may be sprayed into each nostrilof a patient suffering from the common cold. The process is repeatedevery four hours. Within one hour of the first treatment, the patientwill generally report easier breathing through the nose. With two daysof treatments, the patient can usually breathe easily through bothnostrils and reports no sore throat.

EXAMPLE 1

Preparation of squalene ozonide

Squalene is ozonized by preparing a solution of 10 g squalene in 100 mlhexane. Ozone gas (4% in oxygen, from a corona discharge ozonegenerator), is bubbled through this solution via a glass sparger at therate of 5000 cc/min. The reaction is exothermic, and the reactiontemperature is kept within the range of 0° C. to 35° C., preferably 20°C. to more preferably, 22° C. to 24° C., using a cool water bath. Theresulting product is the ozonide of beta carotene, and has a 98% weightgain over squalene.

EXAMPLE 2

Preparation of linalool ozonide

The ozonide of linalool is prepared by bubbling ozone (4% in oxygen,from a corona discharge ozone generator) through 100 ml neat linaloolvia a glass sparger. The reaction is exothermic, and the reactiontemperature is kept within the range of 0° C. to 35° C., preferably 20°C. to 25° C., and more preferably, 22° C. to 24° C., using a cool waterbath. The resulting product is the ozonide of linalool, and has a 31%weight gain over linalool.

EXAMPLE 3

Preparation of Geraniol Ozonide

The ozonide of linalyl acetate was prepared by bubbling ozone (4% inoxygen, from a corona discharge ozone generator) through 5 ml neatgeraniol at the rate of 5000 cc/min. The reaction mixture was cooled ina water bath, and after 20 minutes, the evolution of heat ceased,indicating completion of the ozonation process. The resulting materialhad no odor, and was soluble in polyethylene glycol (600 m.w.),isopropyl myristate, and mineral oil.

EXAMPLE 4

Primary skin irritation test of ozonide of linalool

Six healthy New Zealand White rabbits were tested for skin irritation.Approximately four hours prior to application of the ozonide sample, thebacks of the animals were clipped free of fur. Each rabbit receivedepidermal abrasions with a sterile needle at one test site while theskin at another test site remained intact. A 1.0% solution of linaloolozonide in isopropyl myristate was prepared. A 0.5 ml portion of thetest solution was applied to each site by introduction under a doublegauze layer to an area of skin approximately 1" square. The patches werecovered with a nonreactive tape and the entire test site was wrappedwith a binder. After 24 hours, the binders, tape, test material residuewas removed with 70% isopropyl alcohol. An evaluation was also made at72 hours after application. The reactions were scored according to themethods described in the Federal Hazardous Substances Act. The testsolution had a Primary Irritation Index (PII) of 1.0. According to FHSAregulations, a material with a PII of less than 5.00 is generally notconsidered a primary irritant to the skin.

EXAMPLE 5

Ocular irritation test in the rabbit of the ozonide of linalool

Six healthy New Zealand White rabbits were selected for study. Therabbits' eyes were judged free of irritation prior to the study byexamining with a pen light and under UV light after installation of 2%fluorescein stain. A 1% solution of the ozonide of linalool was preparedin isopropyl myristate. A 0.1 ml portion of this test solution wasinstilled into the lower conjunctival sac of one eye of each rabbit. Thelids were held closed for one second. The opposite eye of each rabbitreceived 0.1 ml of the isopropyl myristate, as control. Eyes wereexamined and the ocular reaction scored according to the "IllustratedGuide for Grading Eye Irritation by Hazardous Substances" (Appendix 1).At 24, 48, and 72 hours post dosing, the eyes were examined with a penlight and reexamined with UV light following fluorescein staining of thecornea. Under the conditions of this test, the test solution wasconsidered a non-irritant to ocular tissues of the rabbit.

EXAMPLE 6

An injectable composition for use in treatment of AIDS

    ______________________________________                                        250 mg/ml   ozonide of geraniol from Example 3                                balance     oil/water emulsion (soybean) with 0.1%                                        lecithin                                                          ______________________________________                                    

EXAMPLE 7

A vaginal suppository for treatment of condylomata acuminata

    ______________________________________                                        2% w/v      Ozonide of geraniol from Example 3                                Balance     Hydrogenated vegetable oil base                                   ______________________________________                                    

EXAMPLE 8

A topical gel effective against warts

    ______________________________________                                         1% w/v         Ozonide of linalool                                           60% w/v         Carbomer 934                                                   1% w/v         Disodium edetate                                              10% w/v         Glycerin                                                      Balance         propylene glycol, 600 m.w.                                    ______________________________________                                    

EXAMPLE 9

A topical cream effective against chicken pox, herpes simplex and otherviral lesions

    ______________________________________                                        2.5% w/v         Ozonide of linalool                                          48% w/v          Propylene glycol                                             30% w/v          Propyl paraben                                               5% w/v           Polysorbate 60                                               10% w/v          Glyceryl monostearate                                        Balance          Mineral oil                                                  ______________________________________                                    

EXAMPLE 10

A nasal inhalant effective against upper respiratory infections

    ______________________________________                                        1 mg/ml           ozonide of citral                                           balance           epal                                                        ______________________________________                                    

EXAMPLE 11

Test of restoration of immune cell levels in an AIDS patient

A patient testing positive for the presence of HIV antibodies anddiagnosed with AIDS was variously treated with the composition ofExample 6 for a period of 99 days. On days 0 through 6, the patientreceived daily intravenous injections of 4.0 ml of the composition ofExample 6. On days 7 to 19, the patient was treated a.q.i.v. with thesame composition. From days 20 through 44, the patient received notreatment. The patient received daily intramuscular injections from days45 through 77. An immunodeficiency screening was performed on days 7,20, 45, and 78. The results, expressed in cells/cmm, are shown in Table2.

                  TABLE 2                                                         ______________________________________                                                 Day 0  Day 7   Day 20  Day 45                                                                              Day 78                                  ______________________________________                                        Total WBC  3700     4300    6600  4900  5300                                  Total lymphs                                                                             1005     2182    2406  1633  1296                                  Total T lymphs                                                                           834      1724    2213  1486  1102                                  Supp-Cytox T8                                                                            381       851     914   702   635                                  Help-Inducer T4                                                                          392       873    1155   702   414                                  ______________________________________                                    

The results show that intra-venous injection of the composition ofExample 6 increased the levels of all types of cells screened. Thesecell levels decreased during the period of no treatment, and remainedrelatively stable during the period of intra-muscular treatment.

It is believed that intra-venous systemic injection in the mannerdescribed in Example 11 is effective in the treatment of other viralinfections as well.

EXAMPLE 12

In vitro anti-viral assay of the ozonide of linalool

A culture of SV-40 is grown in African Green Monkey (AGM) cells. Theculture is harvested in sterile saline. The titer of SV-40 in thesuspension is determined by Standard Plate Count Method in AGM cells. Aworking suspension of SV-40 with a titer of approximately 1.0×10⁷ plaqueforming units (PFUs)/0.1 ml is then prepared. Four aliquots of 1 ml eachof test solution containing 2.0% ozonide of linalool are removed andplaced in separate sterile screw-capped tubes. Each sample is inoculatedwith 0.1 ml of the working suspension of SV-40 to yield a finalconcentration of approximately 1×10⁶ PFUs/1 ml of the product. Thesamples are stored at 20°-25° C. for a total of 28 days. Samples areselected at 7 day intervals to determine the number of viable PFUspresent. A control with uninoculated solution is also stored withsamples selected at the same intervals. At 7 days, and all subsequentsample selections, there are less than 10 PFUs present. No PFUs arepresent in any control sample.

EXAMPLE 13

Preparation of blood products free of viral activity

Blood obtained from a donor is mixed with 0.5g of ozonide of geraniolfrom Example 3 per unit (500 ml) of blood. The blood is then processedin the normal manner. The resulting blood products are free ofdetectable HIV or other viral activity using standard viral assays.

EXAMPLE 14

Test for efficacy of treatment of chicken pox

A small dose (approximately 25 1) of the composition of Example 9 istopically applied to each lesion on the left side of a child sufferingfrom chicken pox. Lesions on the right side are treated with thecomposition lacking in active ingredient. Within 24 hours, the lesionson the child's left side are significantly reduced with little or noself-induced trauma from scratching. The lesions on the child's rightside are unchanged in size, and show the effects of trauma fromscratching.

In a manner similar to that employed in Example 15, other viral lesions,such as common warts and herpes lesions may be treated by topicalapplication of a terpene ozonide in a pharmaceutically acceptablecarrier or excipient.

EXAMPLE 15

Test for efficacy of treatment of condylomata acuminata

A 5 ml suppository with the composition of Example 4 is administeredintra-vaginally to one group of patients suffering from condylomataacuminata. A second group of such patients receive a suppository withoutthe active ingredient of Example 4. A third group receives cryogenictreatment of the affected area, a commonly used treatment forcondylomata acuminata. The average size of the lesions in each group isapproximately 2 cm². Within seven days, the patients of the first grouphave reduced reddening of the vagina and within 15 days, colposcopy doesnot reveal papilloma viruses. In the second group of patients, thelesions are unchanged after 15 days. Patients in the third group have nocondylomata lesions immediately after treatment, however, these patientscontinue to complain of pain and bleeding for up to 30 days after theprocedure is performed.

EXAMPLE 16

Treatment of Rheumatoid Arthritis

It has been theorized that rheumatoid arthritis is caused by a viralagent. The antiviral ozonides of the present invention are believed tobe efficacious in treatment of this disease. Thus, a 20% oralpreparation comprising capsules containing citral ozonide in mediumchain triglyceride (MCT) is prepared and is taken twice daily by apatient suffering from rheumatoid arthritis. Each dose delivers 400 mgactive ingredient to the 60 kg patient. After 1 week, the ANA of thepatient has dropped from approximately 2500 to 100, indicating remissionof the disease. Similar treatment is effective against psoriasis.

I claim:
 1. A method of treating a viral infection on an exposed surfaceof a mammal, comprising the topical application to said exposed surfaceof a pharmacologically antiviral effective amount of an ozonide of aterpene in a pharmaceutically acceptable carrier or excipient.
 2. Themethod of claim 1, wherein said terpene is selected from the groupconsisting of: limonene, citronella, alpha-carotene, beta-carotene,Vitamin A, linalool, linalyl acetate, squalene, geraniol, alpha-pinene,loganin, cymene, farnesanes, eudesmanes, acoranes, cedranes,chamigranes, caryophyllanes, illudanes, humulenes, himachalenes,longifolanes, perhydroazulenes, quaianes, quaianolides, germacranes,labdane, clerodanes, abietic acid, phyllocladene, giberellins,ophiobolin A, retigeranic acid, gasgardic acid, lanosterol, euphol,oleanane, ursane, lupeol, hydroxyphopanone, lupanes, hopanes,B-selinene, zingibene, camphene, sabinene, ocimene, myrcene, nerol,citral A, citral B, farnesol, bisabolene, phytol and cecropia juvenilehormone.
 3. The method of claim 1, wherein the exposed surface is theskin of the mammal.
 4. A method of treating common warts by topicallyadministering an ozonide of a terpene according to the method of claim1.